A construction method, bacterial strain and application of Aeromonas verkirea attenuated strain
A technique for the construction of Aeromonas verdeii, which is applied in the field of construction of attenuated strains of Aeromonas verdeii, which can solve problems such as drug residues and environmental pollution, and achieve the effect of reduced toxicity and good attenuation effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0051] Example 1: Construction of Aeromonas vernerii exsA knockout strain using homologous double exchange technology
[0052] 1. Design primers and amplify homology arm fragments
[0053] Without affecting the exsA upstream gene and downstream gene coding, this application selects the sense strand of the DNA sequence 2009697-2010213 including the exsA start codon, with a total of 516 bp nucleotides as the knockout fragment. Design primers exsA-F for its upstream homology arm and downstream homology arm respectively 1 / R 1 and exsA-F 2 / R 2 , and a restriction endonuclease BstXI restriction site was added to the primer, and the recognition sequences of the restriction endonucleases KpnI and SacI required for double digestion of the vector were added to the middle sequence of the BstXI enzyme recognition site. Wherein, the DNA sequence of the upstream homology arm is shown in SEQ ID NO.4, and the DNA sequence of the downstream homology arm is shown in SEQ ID NO.5. The desi...
Embodiment 2
[0069] Example 2: Virulence detection of Aeromonas verkisii exsA knockout strain
[0070] 1. Growth curve determination
[0071] image 3 The growth curves of C4 wild type and exsA knockout type of Aeromonas verkisei under the same conditions. As shown in the figure, compared with C4 wild type, C4-ΔexsA had no significant difference in the growth rate, reproductive ability, perception and absorption of external nutrients. It indicated that the exsA gene had no obvious effect on the growth of Aeromonas verkistan.
[0072] 2. Detection of C4-ΔexsA on tilapia half-lethal concentration (LD50)
[0073] The virus challenge experiment was carried out on tilapia. In the experiment of this application, the method of intraperitoneal injection of tilapia was used to challenge the virus. Divide tilapia into experimental group and control group, with at least 80 fish in each group. The tilapia of the experimental group were injected with C4-ΔexsA of Aeromonas verkisii, while the tila...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com